8- to 12-week old male EAAT3−/− mice and their wild-type CD1 littermates were used in this study. The EAAT3 knockout mice whose exon 1 of the EAAT3 gene was disrupted by a neomycin resistance cassette were confirmed to have no EAAT3 protein [9 (link)–11 (link)]. These mice were backcrossed with wild-type CD-1 wild-type mice for more than 10 generations to produce an EAAT3−/− mouse line before our study. They were also backcrossed with wild-type CD1 mice at least once every 8 generations to prevent genetic drift as recommended from the Banbury Conference [12 (link)]. The CD1 wild-type mice used for backcrossing were from Charles River Laboratories (Wilmington, MA, USA). The CD1 wild-type mice used in this study are colonies generated from the littermates of EAAT3−/− mice during the backcrossing of every 8 generations.
Cd 1 wild type mice
Manufactured by Charles River Laboratories
Sourced in United States, Germany
CD-1 wild-type mice are a commonly used outbred mouse strain. They are genetically diverse and can be used as general purpose research models across a wide range of applications.
Automatically generated - may contain errors
Lab products found in correlation
Hank s balanced salt solution, by Thermo Fisher Scientific (3 mentions)
Collagenase type 2, by Worthington (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Progesterone, by Merck Group (2 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Neurobasal, by Thermo Fisher Scientific (1 mentions)
Laminin coated, by Thermo Fisher Scientific (1 mentions)
Poly ornithine, by Merck Group (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Plastic tissue culture flasks, by Corning (1 mentions)
75 cm2 plastic tissue culture flasks, by Corning (1 mentions)
Estradiol, by Merck Group (1 mentions)
Wild type c57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
Mouse monoclonal anti flag m2 antibody, by Merck Group (1 mentions)
19 protocols using cd 1 wild type mice
1
EAAT3 Knockout Mouse Characterization
2
Digit Regeneration in Adult Mice
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Adult 18- to 20-month-old male and female CD1 wild-type mice were purchased from Charles River (Wilmington, MA, United States). Mice were anesthetized with 1–5% isoflurane gas with continuous inhalation. The second and fourth digits of both hind limbs were amputated at the P3 distal level as described previously (Fernando et al., 2011 (link); Sammarco et al., 2014 (link); Busse et al., 2019 ), and regenerating digits were collected at day 42 for analysis. The third digit was used as an unamputated control.
3
Isolation and Culture of Spinal Cord Motoneurons
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CD1 wild-type mice were directly purchased from Charles River (Sulzfeld, Germany). Primary spinal cord motoneurons from mice were cultured essentially as described previously (25 (link)). Briefly, spinal cords of E12.5 mouse embryos were dissected and collected in 1.5-ml tubes containing Hank's Balanced Salt Solution, Life Technologies GmbH, Darmstadt Germany After trypsin treatment and trituration, the suspension was enriched for p75NTR-positive cells via an antibody-panning procedure (25 (link)). After washing and depolarization to remove the cells from the panning dish, the cells were collected via centrifugation at 400g for 5 min and resuspended in full medium (Neurobasal (Invitrogen), 1× GlutaMax (Invitrogen), supplemented with 1× B27 (Invitrogen), 2% horse serum (Linaris, Dossenheim, Germany)). After counting with a hemocytometer, 7 × 105 cells were plated on poly-ornithine- (Sigma) and laminin-coated (Invitrogen) dishes (six-well format, Delta surface, Nunc/ Thermo Fisher Scientific Inc., Waltham, MA) and fed with full medium supplemented with 5 ng/ml BDNF. 40% of the medium was replaced after 24 h and every second day after that. In total, cells from ∼140 CD1 wild-type embryos were used in three experiments. After 7 days in culture, cells were washed three times with PBS and harvested in 0.5 ml of lysis buffer (4% SDS, 0.1 m DTT, 10 mm Hepes, pH 8.0) per well.
4
Isolation of Neonatal Mouse Cardiomyocytes
5
Isolation of Neonatal Cardiomyocytes from Mice
6
Assessing Uterine Progesterone Responses in Mice
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All experimental procedures involving mice followed a protocol approved by the Washington University in St. Louis Institutional Animal Care and Use Committee (Protocol Number 20160227). CD1 wild-type mice (Charles River, Saint Louis, MO, United States) were maintained on a 12-h light:12-h dark cycle. To assess uterine progesterone responses, 6-week-old CD1 mice were bilaterally ovariectomized, allowed to rest for 2 weeks to allow the endogenous ovarian-derived steroid hormones to dissipate, and then subcutaneously injected with 100 μl of sesame oil (vehicle control) or 1 mg of progesterone (Sigma-Aldrich) in 100 μl of sesame oil. Six hours later, mice were euthanized, uterine tissues were collected, and RNA was isolated and processed for qRT-PCR.
7
CD1 Mouse Breeding Protocol
8
Digit Regeneration in CD1 Mice
9
Axin2-Deficient Mice: Comprehensive Protocol
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All experiments with animals were performed following Stanford University Animal Care and Use Committee guidelines. All research on animals has been approved by Stanford APLAC, protocol #9999, following approved guidelines by the Stanford University's Institutional Review Board. CD–1 wild type mice were purchased from Charles River Laboratories Inc. Axin2(LacZ/LacZ) homozygotic mice, were genotyped as previously described [19 (link)]. Herein, these mice are referred as to Axin2-/-. Animals were housed in light- and temperature-controlled rooms and were given food and water ad libitum.
10
Uterine Estrogen and Progesterone Responses
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All experimental procedures with mice followed a protocol approved by the Washington University in St. Louis Institutional Animal Care and Use Committee (Protocol Number: 20191079). CD1 wild-type mice (Charles River, Saint Louis, MO) were maintained on a 12-h light: 12-h dark cycle. Sexually mature (8-week-old) CD1 females were mated to fertile wild-type males, and copulation was confirmed by the presence of vaginal plug the following morning, designated as 1 day post-coital (dpc). Mice were euthanized, and uteri were collected on 1, 2, 3, 4, 5, and 6 dpc. To determine the uterine estrogen or progesterone responses, 6-week-old CD1 mice (n = 5 mice per group) were bilaterally ovariectomized, rested for 2 weeks to allow the endogenous ovarian-derived steroid hormones to dissipate, and then subcutaneously injected with 100-μl sesame oil (vehicle control), 1-mg progesterone, or 100-ng estradiol (Sigma-Aldrich, Saint Louis, MO, USA) in 100-μl sesame oil. Six hours later, the mice were euthanized, uterine tissues were collected and fixed in 4% paraformaldehyde, and RNA was isolated and processed for qRT-PCR [17 (link)].
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